Dolichyl phosphates have been described as carriers of carbohydrate units in the biosynthesis of glycoproteins in animal tissues. The investigations proposed are designed to isolate and characterize dolichyl pyrophosphate synthetase and dolichol kinase. The mechanism of the formation of these unique polyprenol products will be of particular interest. In vitro studies will be made with several animal tissues but most of studies will use bovine liver. The kinase and synthetase will be solubilized from one or more of these tissues by extracting isolated membranes with detergents, urea, salt and/or nonpolar solvents. These enzymes will be purified by a combination of fractionation techniques. The synthetase specificity will be investigated with respect to the stereochemistry and chain length of the allylic pyrophosphate substrates. Purified dolichol kinase will also be studied with respect to its substrate specificity. Comparison of other reaction requirements with those described for the analogous bacterial (undecaprenol) kinase will be of particular interest. Other important factors, particularly phospholipid requirements, will also be studied for both enzymes. The enzymically produced polyprenyl pyrophosphates will be characterized after hydrolysis to the free polyprenols. These alcohols will be purified by reverse TLC and HPLC and characterized with respect to both chain length and stereochemistry.